Automated system for two-dimensional electrophoresis

Abstract

The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

What is claimed is:  
     
       1. An automated computer-controlled method for isolating one or more selected proteins present in a two-dimensional array prepared by two-dimensional electrophoresis in a gel, the method comprising: 
       a) laying only the gel smoothly on a scanning platform in the absence of an attached backing material;  
       b) scanning the gel to yield a digitized image;  
       c) inputting said image into an image processor for identifying a plurality of protein-containing regions in the gel;  
       d) excising said protein-containing regions from said gel by means of a computer-controlled excision device on the basis of coordinates supplied by said image processor, said excision device being moved by a computer-controlled means to cut, retrieve and expel a plug of the gel; and  
       e) depositing said protein-containing regions in a plurality of vessels.  
     
     
       2. An automated, computer controlled method for analysis of species in a biological sample by subjecting the sample to two dimension electphoresis comprising the steps of: 
       (i) separating the species by applying the contents of the sample to an electrophoresis gel;  
       (ii) laying only the gel on an imaging platform, the gel being loose and free of any support other than the imaging platform that would interfere with detection of the species;  
       (iii) imaging the resultant array of species;  
       (iv) selecting a point of the array, as containing species for analysis;  
       (v) removing a portion of the gel at the selected point;  
       (vi) analyzing the contents of the removed portion of the gel; and  
       (vii) repeating steps (iv), (v) and (vi), for another point of the array, thereby recovering and characterizing individual species.  
     
     
       3. An automated, computer controlled method for analysis of species in a biological sample as set forth in  claim 2 , wherein said separating step comprises computer controlled electrophoresis equipment wherein a computer for said computer control includes a database for storing species data including location and abundance information. 
     
     
       4. An automated, computer controlled method for analysis of species in a biological sample as set forth in  claim 2 , wherein said imaging step includes scanning the gel to produce a digital image of the gel, the digital image including location and abundance information concerning the separated species. 
     
     
       5. An automated, computer controlled method for analysis of species in a biological sample as set forth in  claim 2 , wherein: 
       said imaging step includes generating a computer-readable output corresponding to the imaged array of species; and  
       said selecting step includes generating machine-readable instructions to remove the selected point of the gel.  
     
     
       6. An automated, computer controlled method for analysis of species in a biological sample as set forth in  claim 5 , wherein said removing step includes using the generated machine-readable instructions to cause a spot cutting machine to remove the selected portion of the gel at the selected point. 
     
     
       7. An automated, computer controlled method for analysis of species in a biological sample as set forth in  claim 2 , wherein said analyzing step includes running the species contained within the removed portion of the gel through a mass spectrometer.